LOCUS pPink-HC 7667 bp ds-DNA circular SYN 04-二月-2013
DEFINITION Pichia pastoris integrating vector for high-copy expression in the
PichiaPink(TM) system.
ACCESSION .
VERSION .
KEYWORDS pPink-HC
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT Linearize within PpTRP2 for integration.
FEATURES Location/Qualifiers
source 1..7667
/organism="synthetic DNA construct"
/lab_host="Pichia pastoris"
/mol_type="other DNA"
promoter 2..940
/gene="Pichia pastoris AOX1"
/note="AOX1 promoter"
/note="inducible promoter, regulated by methanol"
/note="color: #ffffff; direction: RIGHT"
misc_feature 941..991
/note="MCS"
/note="multiple cloning site"
/note="color: #99ccff"
terminator 1017..1266
/gene="S. cerevisiae CYC1"
/note="CYC1 terminator"
/note="transcription terminator for CYC1"
/note="color: #ffffff"
promoter 1286..1298
/gene="Pichia pastoris ADE2"
/note="PpADE2 HC promoter"
/note="weak 13-bp promoter fragment that requires high-copy
integration for efficient expression"
/note="color: #ffffff; direction: RIGHT"
CDS 1299..2990
/codon_start=1
/gene="Pichia pastoris ADE2"
/product="phosphoribosylaminoimidazole carboxylase from
Pichia pastoris"
/note="PpADE2"
/note="essential for the biosynthesis of purine
nucleotides, including adenine"
/note="color: #ff7f50"
promoter 3217..3366
/gene="Pichia pastoris TRP2"
/note="PpTRP2 promoter"
/note="promoter for Pichia pastoris TRP2"
/note="color: #ffffff; direction: RIGHT"
CDS 3367..4983
/codon_start=1
/gene="Pichia pastoris TRP2"
/product="anthranilate synthase from Pichia pastoris"
/note="PpTRP2"
/note="catalyzes the initial step of tryptophan
biosynthesis"
/note="color: #ff7f50"
rep_origin complement(5447..6035)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/note="color: #ffff00"
CDS complement(6206..7066)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/note="This feature has 2 segments:
???1:?6206?..?6997?/?#ccffcc
???2:?6998?..?7066?/?#ccffcc?/?signal?sequence
Cleavage site after base 6997"
promoter complement(7067..7171)
/gene="bla"
/note="AmpR promoter"
/note="color: #ffffff; direction: LEFT"
ORIGIN
1 agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag
61 gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt
121 tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc
181 agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta
241 acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta
301 tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg
361 agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct
421 gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg
481 ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt
541 cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct
601 ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct
661 ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact
721 gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat
781 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt
841 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga
901 caacttgaga agatcaaaaa acaactaatt attcgaaacg gaattccgga ccggcatgcc
961 aaggcctcag gtaccggccg gccatttaaa tacaggcccc ttttcctttg tcgatatcat
1021 gtaattagtt atgtcacgct tacattcacg ccctcctccc acatccgctc taaccgaaaa
1081 ggaaggagtt agacaacctg aagtctaggt ccctatttat tttttttaat agttatgtta
1141 gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacaa acgcgtgtac
1201 gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta
1261 atttgcaagc tggatccgcg gccgccttcc aaactctcat ggattctcag gtaataggta
1321 ttctaggagg aggccagcta ggccgaatga ttgttgaggc cgctagcagg ctcaatatca
1381 agaccgtgat tcttgatgat ggtttttcac ctgctaagca cattaatgct gcgcaagacc
1441 acatcgacgg atcattcaaa gatgaggagg ctatcgccaa gttagctgcc aaatgtgatg
1501 ttctcactgt agagattgag catgtcaaca cagatgctct aaagagagtt caagacagaa
1561 ctggaatcaa gatatatcct ttaccagaga caatcgaact aatcaaggat aagtacttgc
1621 aaaaggaaca tttgatcaag cacaacattt cggtgacaaa gtctcagggt atagaatcta
1681 atgaaaaggc gctgcttttg tttggagaag agaatggatt tccatatctg ttgaagtccc
1741 ggactatggc ttatgatgga agaggcaatt ttgtagtgga gtctaaagag gacatcagta
1801 aggcattaga gttcttgaaa gatcgtccat tgtatgccga gaagtttgct ccttttgtta
1861 aagaattagc ggtaatggtt gtgagatcac tggaaggcga agtattctcc tacccaaccg
1921 tagaaactgt gcacaaggac aatatctgtc atattgtgta tgctccggcc agagttaatg
1981 acaccatcca aaagaaagct caaatattag ctgaaaacac tgtgaagact ttcccaggcg
2041 ctggaatctt cggagttgag atgttcctat tgtctgatgg agaacttctt gtaaatgaga
2101 ttgctccaag gccccacaat tctggtcact atacaatcga tgcatgtgta acatctcagt
2161 tcgaagcaca tgtaagagcc ataactggtc tgccaatgcc actagatttc accaaactat
2221 ctacttccaa caccaacgct attatgctca atgttttggg tgctgaaaaa tctcacgggg
2281 aattagagtt ttgtagaaga gccttagaaa cacccggtgc ttctgtatat ctgtacggaa
2341 agaccacccg attggctcgt aagatgggtc atatcaacat aataggatct tccatgttgg
2401 aagcagaaca aaagttagag tacattctag aagaatcaac ccacttacca tccagtactg
2461 tatcagctga cactaaaccg ttggttggag ttatcatggg ttcagactct gatctacctg
2521 tgatttcgaa aggttgcgat attttaaaac agtttggtgt tccattcgaa gttactattg
2581 tctctgctca tagaacacca cagagaatga ccagatatgc ctttgaagcc gctagtagag
2641 gtatcaaggc tatcattgca ggtgctggtg gtgctgctca tcttccagga atggttgctg
2701 ccatgactcc gttgccagtc attggtgttc ctgtcaaggg ctctacgttg gatggtgtag
2761 actcgctaca ctcgattgtc caaatgccta gaggtgttcc tgtggctacg gttgctatca
2821 acaacgccac caatgccgct ctgttggcca tcaggatttt aggtacaatt gaccacaaat
2881 ggcaaaagga aatgtccaag tatatgaatg caatggagac cgaagtgttg gggaaggcat
2941 ccaacttgga atctgaaggg tatgaatcct atttgaagaa tcgtctttga atttagtatt
3001 gttttttaat agatgtatat ataatagtac acgtaactta tctattccat tcataatttt
3061 attttaaagg ttcggtagaa atttgtcctc caaaaagttg gttagagcct ggcagttttg
3121 ataggcatta ttatagattg ggtaatattt accctgcacc tggaggaact ttgcaaagag
3181 cctcatgtgc ggcgcgccag gccataatgg ccaaacggtt tctcaattac tatatactac
3241 taaccattta cctgtagcgt atttcttttc cctcttcgcg aaagctcaag ggcatcttct
3301 tgactcatga aaaatatctg gatttcttct gacagatcat cacccttgag cccaactctc
3361 tagcctatga gtgtaagtga tagtcatctt gcaacagatt attttggaac gcaactaaca
3421 aagcagatac acccttcagc agaatccttt ctggatattg tgaagaatga tcgccaaagt
3481 cacagtcctg agacagttcc taatctttac cccatttaca agttcatcca atcagacttc
3541 ttaacgcctc atctggctta tatcaagctt accaacagtt cagaaactcc cagtccaagt
3601 ttcttgcttg aaagtgcgaa gaatggtgac accgttgaca ggtacacctt tatgggacat
3661 tcccccagaa aaataatcaa gactgggcct ttagagggtg ctgaagttga ccccttggtg
3721 cttctggaaa aagaactgaa gggcaccaga caagcgcaac ttcctggtat tcctcgtcta
3781 agtggtggtg ccataggata catctcgtac gattgtatta agtactttga accaaaaact
3841 gaaagaaaac tgaaagatgt tttgcaactt ccggaagcag ctttgatgtt gttcgacacg
3901 atcgtggctt ttgacaatgt ttatcaaaga ttccaggtaa ttggaaacgt ttctctatcc
3961 gttgatgact cggacgaagc tattcttgag aaatattata agacaagaga agaagtggaa
4021 aagatcagta aagtggtatt tgacaataaa actgttccct actatgaaca gaaagatatt
4081 attcaaggcc aaacgttcac ctctaatatt ggtcaggaag ggtatgaaaa ccatgttcgc
4141 aagctgaaag aacatattct gaaaggagac atcttccaag ctgttccctc tcaaagggta
4201 gccaggccga cctcattgca ccctttcaac atctatcgtc atttgagaac tgtcaatcct
4261 tctccataca tgttctatat tgactatcta gacttccaag ttgttggtgc ttcacctgaa
4321 ttactagtta aatccgacaa caacaacaaa atcatcacac atcctattgc tggaactctt
4381 cccagaggta aaactatcga agaggacgac aattatgcta agcaattgaa gtcgtctttg
4441 aaagacaggg ccgagcacgt catgctggta gatttggcca gaaatgatat taaccgtgtg
4501 tgtgagccca ccagtaccac ggttgatcgt ttattgactg tggagagatt ttctcatgtg
4561 atgcatcttg tgtcagaagt cagtggaaca ttgagaccaa acaagactcg cttcgatgct
4621 ttcagatcca ttttcccagc aggaaccgtc tccggtgctc cgaaggtaag agcaatgcaa
4681 ctcataggag aattggaagg agaaaagaga ggtgtttatg cgggggccgt aggacactgg
4741 tcgtacgatg gaaaatcgat ggacacatgt attgccttaa gaacaatggt cgtcaaggac
4801 ggtgtcgctt accttcaagc cggaggtgga attgtctacg attctgaccc ctatgacgag
4861 tacatcgaaa ccatgaacaa aatgagatcc aacaataaca ccatcttgga ggctgagaaa
4921 atctggaccg ataggttggc cagagacgag aatcaaagtg aatccgaaga aaacgatcaa
4981 tgaacggagg acgtaagtag gaatttatgg tttggccata atggcctagc ttggcgtaat
5041 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
5101 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
5161 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
5221 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
5281 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
5341 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
5401 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
5461 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
5521 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
5581 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
5641 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
5701 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
5761 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
5821 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
5881 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
5941 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
6001 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
6061 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
6121 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
6181 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
6241 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
6301 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
6361 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
6421 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
6481 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
6541 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
6601 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
6661 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
6721 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
6781 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
6841 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
6901 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
6961 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
7021 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
7081 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
7141 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
7201 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
7261 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
7321 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
7381 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
7441 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
7501 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
7561 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
7621 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattg
//